The people of the Aberjona Basin live in a heavily contaminated watershed and apprehend real and present danger from exposure to chemicals emanating from two Superfund Sites and many other specific sites designated by the Massachusetts environmental authorities. We propose to initiate within this Program a long term study to discover the nature and, if possible, the causes of genetic change in the humans inhabiting the Aberjona Basin. We will strive in particular, to discover if any of the chemicals associated with the identified chemical dump sites are contributing to genetic change in the population. If we succeed our methods will be applicable to the study of any human population. Like our collaborators in the hydrologic engineering projects our proposal involves development of a general theory, creation of new analytical tools to test the theory and the use of these new tools in the context of the Aberjona Basin. Since every mutagen (studied in phage, bacteria, rodent and human cells) has produced a unique set of mutations with regard to kind and position our theory is that knowledge of such sets or mutational spectra in human tissue samples will yield invaluable information leading to the diagnosis of the primary cause(s) of these mutations. We specifically propose to define the kind and frequency of nuclear and mitochondrial mutations in blood samples of five multigenerational families residents in the Aberjona Valley. We will use this initial sample to test the statistical null hypotheses that (a) there are nor nonrandom features in the mutational spectra of ordinary citizens [Useful mutational spectral exist.] (b) There are no significant differences among the mutational spectra of unrelated citizens. [There is a primary cause of mutation in all people]. (c) There are no significant differences among the mutational spectra of members of the same family [There is a primary cause of mutation related to both genetic make up and environmental exposure]. (d) There are no significant differences between the mutational spectra for any individual and that produced by treatment of human B cells with suspect surface and ground water contaminants found in Aberjona water and sediments. Specifically - raw mixed sedimentary mutagens, chromium compounds, arsenicals, organic solvents and other such pure mutagenic compounds as may be identified in the work of Professor Hemond will have their mutational spectra determined. While the initial population study involves five multigenerational families living in the Aberjona, a significant amount of effort must still be invested in development and calibration of the general mensuration technology. Certain specific improvements in separation technology complementary to improvements in DNA amplification fidelity described in Project 0002 are made which are essential to our success.